Details

Autor(en) / Beteiligte

• Kapoor, Aastha
• Chen, Carolyn G.
• Iozzo, Renato V.

Titel

A simplified aortic ring assay: A useful ex vivo method to assess biochemical and functional parameters of angiogenesis

Ist Teil von

• Matrix biology plus, 2020-05, Vol.6-7, p.100025-100025, Article 100025

Ort / Verlag

Elsevier B.V

Erscheinungsjahr

2020

Link zum Volltext

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• We present a simplified method for conducting aortic ring assays which yields robust sprouting and high reproducibility targeted towards matrix biologists studying angiogenesis and extracellular matrix signaling. Main adjustments from previously established protocols include embedding aortic rings between two layers of 3D type I collagen matrix and supplementing with vascular endothelial media. We also introduce a concise and effective staining protocol for obtaining high-resolution images of intracellular and extracellular matrix proteins along with a more accurate protocol to quantify angiogenesis. Importantly, we present a novel method to perform biochemical analyses of vessel sprouting without contamination from the aortic ring itself. Overall, our refined method enables detection of low abundance and phosphorylated proteins and provides a straightforward ex vivo angiogenic assay that can be easily reproduced by those in the matrix biology field. •We report a simplified ex vivo aortic ring assay with enhanced sprouting.•We use a two-layered 3D collagen matrix to encapsulate aortic rings.•We obtain high-resolution images of intracellular and extracellular matrix proteins.•We achieve reproducible biochemical and immunological analyses of aortic rings.